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| author | Bonface | 2024-02-09 09:41:28 -0600 |
|---|---|---|
| committer | Munyoki Kilyungi | 2024-08-09 13:30:43 +0300 |
| commit | d029d5d7f8ead1f1de8d318045004a4a6f68f5fb (patch) | |
| tree | 33c7ff40e3f953d030ed08f468f7afb1dfcba9e6 /general/datasets/TIGEM_hg38_ret_rna_seq_0916 | |
| parent | 769ff7825f5d8d36d541e90534c07f1985899973 (diff) | |
| download | gn-docs-d029d5d7f8ead1f1de8d318045004a4a6f68f5fb.tar.gz | |
Update dataset RTF Files.
Diffstat (limited to 'general/datasets/TIGEM_hg38_ret_rna_seq_0916')
5 files changed, 15 insertions, 0 deletions
diff --git a/general/datasets/TIGEM_hg38_ret_rna_seq_0916/acknowledgment.rtf b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/acknowledgment.rtf new file mode 100644 index 0000000..54bc7ff --- /dev/null +++ b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/acknowledgment.rtf @@ -0,0 +1 @@ +<p>The authors are grateful to Mohit Parekh from the Fondazione Banca degli Occhi del Veneto (FBOV) for the collection of human retina samples. The authors would also like to thank Manuela Dionisi (technician) and Vincenzo Nigro (Head) of the Next Generation Sequencing Facility (TIGEM). This work was supported by FondazioneTelethon Grant (TGM11SB2) to SB and (TGM11SB1) toDdB and to the EU FP7 Radiant project (305626) to DdB.</p>
diff --git a/general/datasets/TIGEM_hg38_ret_rna_seq_0916/cases.rtf b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/cases.rtf new file mode 100644 index 0000000..71f6c2f --- /dev/null +++ b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/cases.rtf @@ -0,0 +1,3 @@ +<h3>Human retina sample collection</h3>
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+<p>Retina samples were collected at Fondazione Banca degli Occhi del Veneto (FBOV) from 50 different donors for cornea transplantation in compliance with the tenets of the Declaration of Helsinki and after an informed consent allowing the use of tissues for research purposes was signed by the donor's next of kin (for a description of donors Supplementary Table S1). Each harvested tissue was accompanied with the FBOV progressive number and with details on the age and gender of donor, the cause of death and the total post-mortem time (T). To limit the possible effects of post-mortem time on RNA integrity and transcriptomic profiles, retinal tissues were isolated only from eye bulbs with a total post-mortem interval (T) ≤ 26 h. The average post-mortem time of the samples was 20.5 h (ranging from 6 to 26 h). For the same reason, bulbs deriving from multi-organ donors were excluded from the analysis. In order to limit cross-contamination with adjacent tissues, we established a protocol for the dissection of the retina from the eye bulbs (<a href="http://nar.oxfordjournals.org/content/early/2016/05/27/nar.gkw486.full#ref-14" id="xref-ref-14-1">14</a>). The dissected retinal tissue was visually inspected to exclude any cross-contamination with the pigmented RPE/choroid and was immediately submerged in RNA Stabilization Reagent (RNA later; QIAGEN).</p>
diff --git a/general/datasets/TIGEM_hg38_ret_rna_seq_0916/experiment-design.rtf b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/experiment-design.rtf new file mode 100644 index 0000000..f9da145 --- /dev/null +++ b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/experiment-design.rtf @@ -0,0 +1,3 @@ +<h3>RNA extraction, library preparation and sequencing</h3>
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+<p>Total RNA was extracted from the 50 human retina samples using the miRNeasy Kit (QIAGEN) according to the manufacturer's instructions. RNA was quantified using a NanoDrop ND-8000 spectrophotometer (NanoDrop Technologies) and the integrity was evaluated using an RNA 6000 Nano chip on a Bioanalyzer (Agilent Technologies). The RNA of the 50 samples had an average RNA integrity number (RIN) of 8.7 (ranging from 7.2 to 9.7). Libraries were prepared according to manufacturer's instructions (TruSeq RNA Sample Preparation kit) with an initial amount of 4 μg of total RNA. Quality control of library templates was performed using a High Sensitivity DNA Assay kit (Agilent Technologies) on a Bioanalyzer (Agilent Technologies). Qubit quantification platform was used to normalize samples for the library preparation (Qubit 2.0 Fluorometer, Life Technologies). Libraries were sequenced via a paired-end chemistry on an Illumina HiSeq1000 platform with an average yield of ∼6 Mb.</p>
diff --git a/general/datasets/TIGEM_hg38_ret_rna_seq_0916/processing.rtf b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/processing.rtf new file mode 100644 index 0000000..7d8d4b3 --- /dev/null +++ b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/processing.rtf @@ -0,0 +1,3 @@ +<h3>Data analysis</h3>
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+<p>The exploratory analysis, whose steps are shown in Figure <a href="http://nar.oxfordjournals.org/content/early/2016/05/27/nar.gkw486.full#F1" id="xref-fig-1-1">1</a>, was carried out with the ‘tuxedo’ software suite (Trapnell <em>et al</em>., 2010) and led to the definition of the Observed Transcriptome (ObsT). The conservative analysis (Figure <a href="http://nar.oxfordjournals.org/content/early/2016/05/27/nar.gkw486.full#F1" id="xref-fig-1-2">1</a>) was carried out by running the RNA-Seq by Expectation-Maximization (RSEM) package (Li & Dewey, 2011) and led to the definition of the RefT.</p>
diff --git a/general/datasets/TIGEM_hg38_ret_rna_seq_0916/summary.rtf b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/summary.rtf new file mode 100644 index 0000000..52d3ca2 --- /dev/null +++ b/general/datasets/TIGEM_hg38_ret_rna_seq_0916/summary.rtf @@ -0,0 +1,5 @@ +<p>Citation: ​ <b>Pinelli</b> M, Carissimo A, Cutillo L, Lai CH, Mutarelli M, Moretti MN, Singh MV, Karali M, Carrella D, Pizzo M, Russo F, Ferrari S, Ponzin D, Angelini C, Banfi S, di Bernardo D (2016) <a data-href="/citations?view_op=view_citation&hl=it&user=5vU8xssAAAAJ&citation_for_view=5vU8xssAAAAJ:9ZlFYXVOiuMC">An atlas of gene expression and gene co-regulation in the human retina</a>. Nucleic acids research 44 (12), 5773-5784</p>
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+<p> </p>
+
+<p>The human retina is a specialized tissue involved in light stimulus transduction. Despite its unique biology, an accurate reference transcriptome is still missing. Here, we performed gene expression analysis (RNA-seq) of 50 retinal samples from non-visually impaired post-mortem donors. We identified novel transcripts with high confidence (Observed Transcriptome (ObsT)) and quantified the expression level of known transcripts (Reference Transcriptome (RefT)). The ObsT included 77 623 transcripts (23 960 genes) covering 137 Mb (35 Mb new transcribed genome). Most of the transcripts (92%) were multi-exonic: 81% with known isoforms, 16% with new isoforms and 3% belonging to new genes. The RefT included 13 792 genes across 94 521 known transcripts. Mitochondrial genes were among the most highly expressed, accounting for about 10% of the reads. Of all the protein-coding genes in Gencode, 65% are expressed in the retina. We exploited inter-individual variability in gene expression to infer a gene co-expression network and to identify genes specifically expressed in photoreceptor cells. We experimentally validated the photoreceptors localization of three genes in human retina that had not been previously reported. RNA-seq data and the gene co-expression network are available online (<a href="http://retina.tigem.it/">http://retina.tigem.it</a>).</p>
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